Effects of a SETD2 mutation on DNA damage response in renal cell carcinoma
Open Access
- Author:
- Peskin, Jessica
- Area of Honors:
- Biology (Abington)
- Degree:
- Bachelor of Science
- Document Type:
- Thesis
- Thesis Supervisors:
- Thomas Michael Mcguire, III, Thesis Supervisor
David Ruth, Thesis Honors Advisor - Keywords:
- SETD2
Renal cell carcinoma
DNA damage and repair - Abstract:
- SETD2 is a histone methyltransferase that trimethylates histone H3K36. Normally, SETD2 functions as a tumor suppressor gene and is involved in homologous recombination (HR) DNA repair of double stranded breaks (DSBs), however mutations in SETD2 arise in about 10% of renal cell carcinomas (RCC). Labs showed that there are HR defects associated with SETD2 loss. We aimed to characterize these HR defects in Setd2-mutated vs WT RENCA cells. We aimed to determine if HR deficiency is caused by a lack of DSB detection or a lack of repair after a DSB is detected. Nuclear foci formation at DSBs marks their detection and early repair and could thus be used to determine if DSBs were detected. Western blotting was used to determine DNA damage protein activation (DSB repair). We utilized the HR-GFP plasmid to determine if DSBs are repaired as well. We hypothesized Setd2-mutant cells would be more sensitive to PARP inhibitors and evaluated that using cell viability. We show that Setd2 mutant cells do not die upon buildup of DNA damage, which indicates an impaired DNA damage repair system. This could be from a lack of p53 expression found in Setd2 mutant cells. Furthermore, Setd2 mutant vs WT cell lines showed similar effects to Olaparib PARP inhibitor, indicating no therapeutic susceptibility. Other DNA damaging agents like Etoposide also indicated no therapeutic susceptibility.